Registration Dossier

Diss Factsheets

Administrative data

Key value for chemical safety assessment

Effects on fertility

Description of key information

There is a developmental toxicity study acc. OECD TG 414 available.

According to the ECHA Factsheet (ECHA, 2009) regarding the information requirements for reproductive toxicity, ECHA considers registration dossiers for substances >= 100 t/y (and 1000 t/y) as technically complete if the dossier contains a prenatal developmental toxicity study in accordance with Section 8.7.2 of column 1, Annex IX. In this dossier, a prenatal developmental toxicity gavage study (rat, OECD 414) was conducted in accordance with ECHA Decision TPE-D-2114306081 -68 -01/F Helsinki, 30 July 2015.

Link to relevant study records
Reference
Endpoint:
extended one-generation reproductive toxicity - basic test design (Cohorts 1A, and 1B without extension)
Data waiving:
other justification
Justification for data waiving:
the study does not need to be conducted because a pre-natal developmental toxicity study is available
Reproductive effects observed:
not specified
Effect on fertility: via oral route
Endpoint conclusion:
no study available
Effect on fertility: via inhalation route
Endpoint conclusion:
no study available
Effect on fertility: via dermal route
Endpoint conclusion:
no study available
Additional information

According to the ECHA Factsheet (ECHA, 2009) regarding the information requirements for reproductive toxicity, ECHA considers registration dossiers for substances >= 100 t/y (and 1000 t/y) as technically complete if the dossier contains a testing proposal for a pre-natal developmental toxicity study in accordance with Section 8.7.2 of column 1, Annex IX. In this dossier, a pre-natal developmental toxicity gavage study (rat, gavage, OECD 414) is available. Therefore, a extended one generation reproductive toxicity study (OECD 443) does not need to be conducted.


Short description of key information:
No studies have been performed to explicitly address the question of reproductive effects in animals caused by cyclohexane, 5-isocyanato-1-(isocyanatomethyl)-1,3,3-trimethyl-, homopolymer, acetone oxime-blocked. A data waiver is claimed.

Effects on developmental toxicity

Description of key information

For cyclohexane, 5 -isocyanato-1 -(isocyanatomethyl)-1,3,3 -trimethyl-, homopolymer, acetone oxime-blocked a prenatal developmental toxicity (oral) study according to OECD 414 was conducted in accordance with ECHA Decision TPE-D-2114306081 -68 -01/F Helsinki, 30 July 2015.

Link to relevant study records

Referenceopen allclose all

Endpoint:
developmental toxicity
Type of information:
experimental study
Adequacy of study:
supporting study
Study period:
2015-11-16 to 2015-12-09
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Justification for type of information:
Study was conducted as dose range finding study (DRF) according ECHA Decision TPE-D-2114306081-68-01/F Helsinki, 30 July 2015
Qualifier:
according to guideline
Guideline:
OECD Guideline 414 (Prenatal Developmental Toxicity Study)
Version / remarks:
adopted January 22, 2001
Qualifier:
according to guideline
Guideline:
EU Method B.31 (Prenatal Developmental Toxicity Study)
Version / remarks:
May 30, 2008
GLP compliance:
yes (incl. QA statement)
Limit test:
no
Species:
rat
Strain:
CD-1
Details on test animals or test system and environmental conditions:
TEST ORGANISMS: 
- Species: Rat
- Source: Charles River Laboratories, Research Models and Services, Germany GmbH
- Strain: CD / Crl: CD(SD)
- Age: 60 - 62 days
- body weight: 192.3 to 233.5 g
- Diet: ad libitum, Commercial diet ssniff® R/Z V1324 (ssniff Spezialdiäten GmbH, 59494 Soest, Germany)
- Water: ad libitum
- Acclimatisation period: 5 days
-Housing: Except during the mating period, the dams were kept singly in MAKROLON cages
ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 °C +/- 3° C
- Humidity (%): 55% +/- 15 %
- Illumination: 12 hours artifical fluorescent light and 12 hours dark
- Ventilation rate: between fifteen to twenty air changes per hour.
Route of administration:
oral: gavage
Vehicle:
corn oil
Details on exposure:
ADMINISTRATION: 
- Frequency: once daily, day 6 to 20 of gestation
- Dose volume: 3 ml/kg b.w.
- Dose: 0, 100, 300, 1000 mg/kg/bw
- Animals: 3 female rats/dose
- DOSAGE PREPARATION: The test item formulations were freshly prepared every day.
The test item was suspended in the vehicle to the appropriate concentration and was administered orally at a constant volume once daily from the
6th to the 20th day of gestation. The amount of the test item was daily adjusted to the current body weight of the animal. The control animals
received the vehicle at the same administration volume daily in the same way.
Analytical verification of doses or concentrations:
no
Details on analytical verification of doses or concentrations:
Tests by appropriate analytical methods will be carried out for the main study (prenatal toxicity study in rats by oral administration), to determine the concentration, homogeneity and, if needed, stability of the test item in the formulations.
Details on mating procedure:
Sexually mature ('proved') male rats of the same breed served as partners. The female breeding partners were randomly chosen.
Mating was monogamous: 1 male and 1 female animal were placed together in one cage during the dark period. Each morning a vaginal smear was taken to check for the pres-ence of sperm. If findings were negative, mating was repeated with the same partner. The day on which sperm was found was considered as the day of conception (day 0 of pregnancy). This procedure was repeated until sufficient pregnant rats were available for all groups.
Duration of treatment / exposure:
From gestation day 6 until gestation day 20
Frequency of treatment:
Once daily
Duration of test:
On gestation day 21, all rats were euthanized by carbon dioxide (CO2) inhalation and laparotomised.
Dose / conc.:
0 mg/kg bw/day (nominal)
Remarks:
Vehicle control
Dose / conc.:
100 mg/kg bw/day (nominal)
Remarks:
Low dose
Dose / conc.:
300 mg/kg bw/day (nominal)
Remarks:
Intermediate dose
Dose / conc.:
1 000 mg/kg bw/day (nominal)
Remarks:
HIgh dose
No. of animals per sex per dose:
3 female rats/dose , orally dosed with 0, 100, 300 or 1000 mg test item/kg b.w.
Evaluated litters: 2 litters per group
Control animals:
yes, concurrent vehicle
Details on study design:
Dose selection rationale:
The dose levels were selected in agreement with the Sponsor/Monitor and were based on available toxicological data
(LD50 cut-off for the rat, oral: > 2000 mg/kg b.w.).
Maternal examinations:
Dated and signed records of all activities relating to the day to day running and maintenance of the study within the animal units, as well as to the
group observations and examinations outlined in the Study Plan, were recorded in the appropriate documentation. In addition, observations relating
to the individual animals made throughout the study were recorded.

The following observations were made during the course of the study:
Clinical signs
Individual animals were observed daily for any signs of behavioural changes, reaction to treatment, or illness.
Immediately after administration, any signs of illness or reaction to treatment were rec-orded. In case of changes, the animals were observed until thesymptoms disappeared. In addition, animals were checked regularly throughout the working day from 7.00 a.m. to 3.45 p.m.
On Saturdays and Sundays, the animals were checked regularly starting from 7.00 a.m. to 11.00 a.m. with a final check performed at approximately
3.30 p.m.
Dated and signed records of appearance, change and disappearance of clinical signs were maintained on clinical history sheets for individual animals.


Viability
Further checks were made early in the morning and again in the afternoon of each working day to look for dead or moribund animals. This allowed
post mortem examinations to be carried out during the working period of that day. On Saturdays and Sundays, a similar procedure was followed
except that the final check was carried out at approximately midday.
Animals showing signs of abortion or premature delivery would have been sacrificed on the same day. Fetuses obtained this way were examined for
abnormal development, whenever possible. No abortion occurred in the study.


Body weight
The weight of each rat was recorded on day 0 of gestation (the day of detection of a positive mating sign), followed by daily weighings - always at the same time of the day. The body weight gain was also calculated in intervals (i.e. day 0-3, 3-6, 6 9, 9-12, 12-15, 15-18 and 18-21).
Furthermore the net weight change from day 6 is given:

Carcass weight: Carcass weight = Terminal body weight - uterine weight
Net weight change from day 6:
Net weight change from day 6 = Carcass weight - body weight on day 6
These measurements were also used for calculating the daily amount of test item to be administered.


Food and drinking water consumption
The quantity of food consumed by each rat was recorded daily. Food intake per rat (g/rat/day) was calculated using the total amount of food given to and left by each rat in each group on completion of a treatment day.

The relative food consumption (g/kg b.w./day) was calculated using the following formula:
Daily food consumption [g/kg b.w./day]= Total food intake in g / Body weight in g x 1000

Daily monitoring by visual appraisal of the drinking water bottles was maintained throughout the study.

EXAMINATIONS (NECROPSY), Examination of the dams
On gestation day 21, all rats were euthanized by carbon dioxide (CO2) inhalation and laparotomised.
The ovaries and the uteri of the dams were removed and the uteri (in toto) were weighed. Uteri without fetuses were examinated for possible
implantation sites according to SALEWSKI to confirm their pregnancy status.
In order to check for possible drug effects, a dissection with macroscopic examination of the internal organs and placentae of the dams was carried
out on the day of scheduled laparotomy or on the day on which the animals were found dead. In case of macroscopical findings, the affected maternal tissues were preserved in 7% buffered formalin for possible future histopathological examinations.

Ovaries and uterine content:
Corpora lutea
- number per dam
- absolute number per group
- mean per group

Implantations
- number per dam
- distributions in the uterine horns
- absolute number per group
- mean per group

Resorptions
- number per dam
- distributions in the uterine horns
- absolute number per group
- mean per group
- mean % per group
- early resorptions < 2 mm
- Late resorptions > 2 mm

Weight of placentae
- individual data per fetus
- mean per litter
- mean per group
- litter mean per group
- litter mean per sex and group

Weight of fetuses
- individual data per fetus (alive and dead)
- mean per litter
- mean per group
- litter mean per group
- litter mean per sex and group

Fetuses
- number per dam (alive)
- number per dam (dead)
- number of fetuses (alive and dead) per sex and dam
- distribution in the uterine horns
- absolute number of fetuses alive per group
- mean number of fetuses alive per group
- mean % of fetuses alive per group
- male/female ratio (alive and dead)


Runts
- number per dam
- mean per group

Dead fetuses
- number per dam
- mean per group

Malformed fetuses
- individual data per fetus
- type of malformation: number and incidence (%) per group and litter
- number of affected fetuses per group (fetuses affected by several changes were counted as one fetal incidence)

Total malformation rate [%] = malformed fetuses per group / fetuses per group x 100

Fetuses with variations4
- individual data per fetus
- type of variation: number and incidence (%) per group and litter
- Number of affected fetuses per group (fetuses affected by several changes were be counted as one fetal incidence:
Total variation rate [%] = fetuses per group with variations / fetuses per group x 100

Pre-implantation loss (%)
- per dam
- mean per dam
- per group

Pre-implantation loss [%] = corpora lutea - implantations / corpora lutea x 100

Post-implantation loss ( % )
- per dam
- mean per dam
- per group


Post-implantation loss [%] = implantations - living fetuses / implantations x 100



Fetal examinations:
The fetuses were removed and the following examinations performed:
(a) Macroscopic inspection (gross evaluation) of the placentae for example for focal indurations.
(b) The number of fetuses (alive and dead) and placentae (location in uterus and as-signment to the fetus) was determined.
(c) Sex and viability of fetuses were determined. Animals are said to be viable when they are found alive (spontaneous breathing,
spontaneous movement).
(d) Number and size of resorptions were determined.
(e) Corpora lutea in the ovaries, implantations and location of fetuses in the uterus were determined.
(f) Weights of fetuses and weights of the placentae were determined (fetuses were considered as runts if their weight was less than 70% of the mean litter weight).
(g) Fetuses were inspected externally for damages, especially for malformations .
(h) The fetuses were sacrificed by an ether atmosphere.
Statistics:
No statistical analysis was conducted as only 2 animals per group were evaluated.
Indices:
Dose Range Finding
Clinical signs:
no effects observed
Description (incidence and severity):
No test item-related clinical signs were noted at any of the tested dose levels
Mortality:
no mortality observed
Description (incidence):
None of the dams treated with 100, 300 or 1000 mg test item/kg b.w./day or the vehicle (control) died prematurely during the course of the study.
Body weight and weight changes:
no effects observed
Description (incidence and severity):
No test item-related influence was noted on the body weight or the body weight gain.
Food consumption and compound intake (if feeding study):
no effects observed
Description (incidence and severity):
Food consumption was not influenced at any of the tested dose levels.
Food efficiency:
not specified
Water consumption and compound intake (if drinking water study):
no effects observed
Description (incidence and severity):
No test item-related influence was noted for the drinking water consumption at any of the tested dose levels (visual monitoring).
Ophthalmological findings:
not specified
Haematological findings:
not specified
Clinical biochemistry findings:
not specified
Urinalysis findings:
not specified
Behaviour (functional findings):
not specified
Immunological findings:
not specified
Organ weight findings including organ / body weight ratios:
not specified
Gross pathological findings:
no effects observed
Description (incidence and severity):
Macroscopic inspection at laparotomy revealed no test item-related pathological findings.
Neuropathological findings:
not specified
Histopathological findings: non-neoplastic:
not specified
Histopathological findings: neoplastic:
not specified
Description (incidence and severity):
Uterus weight and carcass weight
No test item-related changes in the gravid uterus weight in comparison to the control group were noted in any of the test groups (100, 300 or 1000 mg test item/kg b.w./day).
No test item-related changes were noted for the carcass weight (body weight without the gravid uterus) between the control dams and the test item treated dams (100, 300 or 1000 mg test item/kg b.w./day).

Body weight gain during the treatment period
No test item-related changes were noted for the net body weight gain (without gravid uterus) after the start of treatment from gestation day 6 until gestation day 21 between the control dams and the test item treated dams (100, 300 or 1000 mg test item/kg b.w./day).
Details on results:
No effects was noted
Number of abortions:
no effects observed
Description (incidence and severity):
No test item-related differences for the above mentioned ratios were noted between the dams of the control group and the dams treated with 100, 300 or 1000 mg test item/kg b.w./day
Pre- and post-implantation loss:
no effects observed
Description (incidence and severity):
No test item-related differences for the above mentioned ratios were noted between the dams of the control group and the dams treated with 100, 300 or 1000 mg test item/kg b.w./day
Total litter losses by resorption:
no effects observed
Description (incidence and severity):
No test item-related differences for the above mentioned ratios were noted between the dams of the control group and the dams treated with 100, 300 or 1000 mg test item/kg b.w./day
Early or late resorptions:
no effects observed
Description (incidence and severity):
No test item-related differences for the above mentioned ratios were noted between the dams of the control group and the dams treated with 100, 300 or 1000 mg test item/kg b.w./day
Dead fetuses:
no effects observed
Description (incidence and severity):
No test item-related differences for the above mentioned ratios were noted between the dams of the control group and the dams treated with 100, 300 or 1000 mg test item/kg b.w./day
Changes in pregnancy duration:
not specified
Description (incidence and severity):
Migrated Data from removed field(s)
Field "Effects on pregnancy duration" (Path: ENDPOINT_STUDY_RECORD.DevelopmentalToxicityTeratogenicity.ResultsAndDiscussion.ResultsMaternalAnimals.MaternalDevelopmentalToxicity.EffectsOnPregnancyDuration): not specified
Changes in number of pregnant:
no effects observed
Description (incidence and severity):
No test item-related differences for the above mentioned ratios were noted between the dams of the control group and the dams treated with 100, 300 or 1000 mg test item/kg b.w./day
Details on maternal toxic effects:
no effects
No test item-related influence on the reproductive parameters (number of implantation sites, fetuses, resorptions, pre- and post-implantation indices) were noted between the dams of the control group and the dams of the treatment groups (100 or 300 or 1000 mg test item/kg b.w./day).
see table below "overall remarks"
Dose descriptor:
NOAEL
Effect level:
> 1 000 mg/kg bw/day
Basis for effect level:
other: maternal toxicity
Fetal body weight changes:
no effects observed
Description (incidence and severity):
No dead fetuses were noted in the litters of the dams of the control group and in the litters of the dams treated with 100, 300 or 1000 mg test item/kg b.w./day.
Migrated Data from removed field(s)
Field "Fetal/pup body weight changes" (Path: ENDPOINT_STUDY_RECORD.DevelopmentalToxicityTeratogenicity.ResultsAndDiscussion.ResultsFetuses.FetalPupBodyWeightChanges): no effects observed
Field "Description (incidence and severity)" (Path: ENDPOINT_STUDY_RECORD.DevelopmentalToxicityTeratogenicity.ResultsAndDiscussion.ResultsFetuses.DescriptionIncidenceAndSeverityFetalPupBodyWeightChanges): No test item-related influence on the mean fetal weights per dam was noted at 100, 300 or 1000 mg test irem/kg b.w./day.
Reduction in number of live offspring:
no effects observed
Changes in sex ratio:
effects observed, non-treatment-related
Description (incidence and severity):
No test item-related influence on the ratio of live male fetuses to live female fetuses were noted for all treatment groups (100, 300 or 1000 mg test item/kg b.w./day).
The sex ratios of the pups (male / female) were 1.89 in the control group, 2.00 in the low dose group, 1.60 in the intermediate dose group and 1.08 in the high dose group. The differences between the ratios are considered to be spontaneous and within the variability of the small number of fetuses examined per group.
Changes in litter size and weights:
no effects observed
Description (incidence and severity):
Weight of placentae
No test item-related influence on the mean placental weight per dam was noted at 100, 300 or 1000 mg test item/kg b.w./day.
Changes in postnatal survival:
not examined
External malformations:
no effects observed
Description (incidence and severity):
No macroscopically visible malformations were noted for the fetuses of the control group and the treatment groups (100, 300 or 1000 mg test item/kg b.w./day) during the external examination of the fetuses at laparotomy.
Skeletal malformations:
not examined
Visceral malformations:
no effects observed
Description (incidence and severity):
Variations
No macroscopically visible variations were noted for the fetuses of the control group and the treatment groups (100, 300 or 1000 mg test item/kg b.w./day).
Other effects:
no effects observed
Description (incidence and severity):
Number of runts
No runts were noted in the test item groups (100, 300 or 1000 mg test item/kg b.w./day) or in the control group
Details on embryotoxic / teratogenic effects:
Embryotoxic / teratogenic effects:no effects
Dose descriptor:
NOAEL
Effect level:
1 000 mg/kg bw/day (nominal)
Based on:
test mat.
Basis for effect level:
other: developmental toxicity
Developmental effects observed:
no

 Examination of the dams

Summary of animals examined

Test item

Group 1

Control

Group 2

100 mg/kg

Group 3

300 mg/kg

Group 4

1000 mg/kg

Treated dams

3

3

3

3

Not pregnant dams

0

1

0

0

Dams without viable fetuses

0

0

0

0

Not examined dams

(spare animals)

1

0

1

1

Evaluated litters

2

2

2

2

Summary of animals evaluated

Test item

dose in mg/kg

b.w./day p.o.

Animal nos.

of mated

rats

Animal nos.

of rats with evaluable litters

at laparotomy

Females

not pregnant

(animal nos.)

Dams

with total

implantation loss

(animal nos.)

Reserve

animals, not

examined

(animal nos.)

Control

1 - 3

1, 2

none

none

3

100

4 - 6

4, 6

5

none

none

300

7 - 9

7, 8

none

none

9

1000

10 - 12

10, 11

none

none

12

1.1               Findings

Examination of the dams:

 

Mortality

None of the dams treated with 100, 300 or 1000 mg test item/kg b.w./day or the vehicle (control)died prematurely during the course of the study.

Clinical signs

No test item-related clinical signs were noted at any of the tested dose levels.

Body weight and

body weight gain

 

No test item-related influence was noted on the body weight or the body weight gain.

Food consumption

Food consumption was not influenced at any of the tested dose levels.


Drinking water consumption

No test item-related influence was noted for the drinking water consumption at any of the tested dose levels (visual monitoring).

Necropsy findings

Macroscopic inspection at laparotomy revealed no test item-related pathological findings.

Examination of the fetuses:

No test item-related influence was detected on the prenatal fetal development at 100, 300 or 1000 mg test item/kg b.w./daywith respect to the number of resorptions, number of live fetuses, fetal and placental weights, the values calculated for the post-implantation loss and the sex distribution of fetuses.

Laparotomy revealed no dead fetuses or runts in the test item groups or the control group.


 

Malformations

 

No external malformation was noted.

 

Variations

 

No external variation was noted.

Conclusions:
In conclusion, no embryotoxic properties of the test item were noted up to the highest tested dose of 1000 mg test item/kg b.w./day by oral
administration. Based on the data obtained in this dose-range-finding study, the following dose levels are suggested for LPT Study No. 32704
(Prenatal developmental toxicity study in rats): Group 1: Control, Group 2:  100 mg test item/kg b.w./day, p.o, Group 3: 300 mg test item/kg b.w.
/day, p.o, Group 4: 1000 mg test item/kg b.w./day, p.o
 
Executive summary:

The aim of this dose-range-finding study was to determine the dose levels for a prenatal developmental toxicity study of the test item in pregnant ratwhenadministered orally during the critical period of organogenesis and the fetal development (6th to 20th day of gestation).

 

In this dose-range-finding study for a prenatal developmental toxicity study, the test item was administered orally to female rats at dose levels of 100, 300 or 1000 mg/kg b.w./day from the 6th to 20th day of pregnancy.

 

Under the present test conditions, the no-observed-adverse-effect level (NOAEL) was above 1000 mg test item/kg b.w./day for the dams.

No premature deaths were noted. No test item-related influence was noted on behaviour or external appearance, body weight as well as intake of food and drinking water of the dams at any tested dose level. Necropsy revealed no changes for the dams of any test group.

 

The no-observed-adverse-effect level (NOAEL) for the fetal organism was also above 1000 mg test item/kg b.w./day.

No dead fetuses, no malformations and no test item-related variations were noted at any of the tested dose levels.

In conclusion, no embryotoxic properties of the test item were noted up to the highest tested dose of 1000 mg test item/kg b.w./day by oral administration.

 

Based on the data obtained in this dose-range-finding study, the following dose levels are suggested for LPT Study No. 32704 (Prenatal developmental toxicity study in rats):

Group 1:

Control

Group 2:

 100 mg test item/100/kg b.w./day, p.o

Group 3:

 300 mg test item/kg b.w./day, p.o

Group 4:

1000 mg test item/kg b.w./day, p.o

 

Endpoint:
developmental toxicity
Type of information:
experimental study
Adequacy of study:
key study
Study period:
2016-01-24 to 2016-02-25
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Justification for type of information:
Study was conducted according ECHA Decision TPE-D-2114306081-68-01/F Helsinki, 30 July 2015
Qualifier:
according to guideline
Guideline:
OECD Guideline 414 (Prenatal Developmental Toxicity Study)
Version / remarks:
adopted January 22, 2001
Qualifier:
according to guideline
Guideline:
EU Method B.31 (Prenatal Developmental Toxicity Study)
Version / remarks:
May 30, 2008
GLP compliance:
yes (incl. QA statement)
Limit test:
no
Species:
rat
Strain:
CD-1
Details on test animals or test system and environmental conditions:
TEST ORGANISMS: 
- Species: Rat
- Source: Charles River Laboratories, Research Models and Services, Germany GmbH
- Strain: CD / Crl: CD(SD)
- Age: 59 days
- body weight: 183.6 - 280.8 g
- Diet: ad libitum, Commercial diet ssniff® R/Z V1324 (ssniff Spezialdiäten GmbH, 59494 Soest, Germany)
- Water: ad libitum
- Acclimatisation period: 5 days
-Housing: Except during the mating period, the dams were kept singly in MAKROLON cages
ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 °C +/- 3° C
- Humidity (%): 55% +/- 15 %
- Illumination: 12 hours artifical fluorescent light and 12 hours dark
- Ventilation rate: between fifteen to twenty air changes per hour.
Route of administration:
oral: gavage
Vehicle:
corn oil
Details on exposure:
ADMINISTRATION: 
- Frequency: once daily, day 6 to 20 of gestation
- Dose volume: 3 ml/kg b.w.
- Dose: 0, 100, 300, 1000 mg/kg/bw
- Animals: 25 female rats/group
DOSAGE PREPARATION:
- The test item formulations were freshly prepared every day.
- The test item was suspended in the vehicle to the appropriate concentrations using a magnetic stirrer and was administered orally at a constant volume once daily from the 6th to the 20th day of gestation. Stirring of the formulations was continued until the last animal of the dose group had been dosed.
- The amount of the test item was daily adjusted to the current body weight of the animal.
- The control animals received the vehicle at the same administration volume daily in the same way.
- The male rats for mating remained untreated.
Analytical verification of doses or concentrations:
yes
Remarks:
The analytical method was successfully re-validated by LPT: Analytical Method for the Determination of test item in Formulations with subsequent HPLC UV Detection
Details on analytical verification of doses or concentrations:
For the analysis of the test item-vehicle formulations, samples of approximately 2 mL were taken at the following times and stored at -20°C or colder until analysis at LPT:
At start of dosing
- Analysis of stability and concentration: Immediately after preparation of the formula-tions as well as after 8 and 24 hours storage of formulations at room temperature. (3 samples/test item group), Number of samples: 3 x 3 = 9,
- Homogeneity: At the start of dosing, during (middle) admin-istration and before dosing to the last animal of the test item group. (3 samples/test item group), Number of samples: 3 x 3 = 9,

At the end of the dosing period (at a time when the majority of animals was dosed):
- Analysis of concentration: During treatment with the test item always before administration to the last animal of the dose level group. (1 sample/test item group), Number of samples: 1 x 3 = 3

Sum of all samples: 21
Details on mating procedure:
Sexually mature ('proved') male rats of the same breed served as partners.
The female breeding partners were randomly chosen.
Mating was monogamous: 1 male and 1 female animal were placed together in one cage during the dark period. Each morning a vaginal smear was taken to check for the pres-ence of sperm. If findings were negative, mating was repeated with the same partner. The day on which sperm was found was considered as the day of conception (day 0 of pregnancy). This procedure was repeated until enough pregnant dams were available for all groups.
Rats which did not become pregnant were excluded from the analysis of the results and replaced by other animals.
A post-mortem negative staining according to SALEWSKI was carried out in the replaced animals in order to confirm the non-pregnancy status.
Duration of treatment / exposure:
From gestation day 6 until gestation day 20
Frequency of treatment:
Once daily
Duration of test:
On gestation day 21, all rats were euthanized by carbon dioxide (CO2) inhalation and laparotomised.
Dose / conc.:
0 mg/kg bw/day (nominal)
Remarks:
Vehicle control
Dose / conc.:
100 mg/kg bw/day (nominal)
Remarks:
Low dose
Dose / conc.:
300 mg/kg bw/day (nominal)
Remarks:
Intermediate dose
Dose / conc.:
1 000 mg/kg bw/day (nominal)
Remarks:
HIgh dose
No. of animals per sex per dose:
25 female rats/dose , orally dosed with 0, 100, 300 or 1000 mg test item/kg b.w.
Evaluated litters: 20 litters per group
Control animals:
yes, concurrent vehicle
Details on study design:
The dose levels were selected in agreement with the Sponsor based on the results of a dose-range-finding study for a prenatal developmental toxicity (LPT study no. 32703).
In this dose-range finding study, test item was administered to pregnant female rats at dose levels of 100, 300 or 1000 mg/kg b.w./day orally, by gavage, once daily from gestation day 6 to 20.
No premature deaths were noted.
No test item-related influence was noted on behaviour or external appearance, body weight as well as intake of food and drinking water of the dams at any tested dose level. Necropsy revealed no changes for the dams of any test group.
No embryotoxic properties (no dead fetuses, no malformations and no test item-related variations) were noted at any of the tested dose levels.

Maternal examinations:
Dated and signed records of all activities relating to the day to day running and maintenance of the study within the animal units, as well as to the
group observations and examinations outlined in the Study Plan, were recorded in the appropriate documentation. In addition, observations relating
to the individual animals made throughout the study were recorded.

The following observations were made during the course of the study:
Clinical signs
Individual animals were observed daily for any signs of behavioural changes, reaction to treatment, or illness.
Immediately after administration, any signs of illness or reaction to treatment were rec-orded. In case of changes, the animals were observed until thesymptoms disappeared. In addition, animals were checked regularly throughout the working day from 7.00 a.m. to 3.45 p.m.
On Saturdays and Sundays, the animals were checked regularly starting from 7.00 a.m. to 11.00 a.m. with a final check performed at approximately
3.30 p.m.
Dated and signed records of appearance, change and disappearance of clinical signs were maintained on clinical history sheets for individual animals.


Viability
Further checks were made early in the morning and again in the afternoon of each working day to look for dead or moribund animals. This allowed
post mortem examinations to be carried out during the working period of that day. On Saturdays and Sundays, a similar procedure was followed
except that the final check was carried out at approximately midday.
Animals showing signs of abortion or premature delivery would have been sacrificed on the same day. Fetuses obtained this way were examined for
abnormal development, whenever possible. No abortion occurred in the study.


Body weight
The weight of each rat was recorded on day 0 of gestation (the day of detection of a positive mating sign), followed by daily weighing - always at the same time of the day.
The body weight gain was calculated in intervals (i.e. day 0-3, 3 6, 6-9, 9-12, 12-15, 15-18 and 18-21), for the whole study (gestation day 0 - 21) and for the period after the start of dosing (gestation day 6 to gestation day 21). Furthermore the carcass weight and the net weight gain from day 6 is given.
These values are stated in the report.
These measurements were also used for calculating the daily amount of test item to be administered.

Food and drinking water consumption
The quantity of food consumed by each rat was recorded daily. Food intake per rat (g/rat/day) was calculated using the total amount of food given to and left by each rat in each group on completion of a treatment day.

The relative food consumption (g/kg b.w./day) was calculated using the following formula:
Daily food consumption [g/kg b.w./day]= Total food intake in g / Body weight in g x 1000

Daily monitoring by visual appraisal of the drinking water bottles was maintained throughout the study.

EXAMINATIONS (NECROPSY), Examination of the dams
Dissection technique and evaluation of the animals:
On gestation day 21, the rats were laparotomised under ether narcosis. The ovaries and the uteri of the dams were removed; the gravid uteri (in toto) were weighed. In order to check for possible test item effects, a dissection with macroscopic examination of the internal organs and placentae of the dams was carried out on the day of sacrifice or on the day on which the animals were found dead. In case of macroscopical findings, the affected maternal tissues were preserved in 7% buffered formalin for possible future histopathological examinations.
Organ weights
The following target organs of the dams were weighed: Heart, kidney (2) and liver.
The kidneys were weighed separately and identified as left and right.

Ovaries and uterine content:
Corpora lutea
- number per dam
- absolute number per group
- mean per group

Implantations
- number per dam
- distributions in the uterine horns
- absolute number per group
- mean per group

Resorptions
- number per dam
- distributions in the uterine horns
- absolute number per group
- mean per group
- mean % per group
- early resorptions < 2 mm
- Late resorptions > 2 mm

Weight of placentae
- individual data per fetus
- mean per litter
- mean per group
- mean per sex and group

Weight of fetuses
- individual data per fetus (alive and dead)
- mean per litter
- mean per group
- litter mean per sex and group

Fetuses
- number per dam (alive)
- number per dam (dead)
- number of fetuses (alive and dead) per sex and dam
- distribution in the uterine horns
- absolute number of fetuses alive per group
- mean number of fetuses alive per group
- mean % of fetuses alive per group
- male/female ratio (alive and dead)


Runts
- number per dam
- mean per group

Dead fetuses
- number per dam
- mean per group

Malformed fetuses
- individual data per fetus
- type of malformation: number and incidence (%) per group and litter
- number of affected fetuses per group (fetuses affected by several changes were counted as one fetal incidence)

Total malformation rate [%] = malformed fetuses per group / fetuses per group x 100

Fetuses with variations4
- type of malformation
- individual data per fetus
- number and incidence (%) per group and litter

Total variation rate [%] = fetuses per group with variations / fetuses per group x 100

Fetuses with retardations
- type of retardation
- individual data per fetus
- number and incidence (%) per group and litter

Total retardation rate [%] = fetuses per group with retardations / fetuses per group x 100


Indices of pre-implantation loss and post-implantation loss:

Calculation of group indices (see table 7-1)
Pre implantation
loss [%] = [Corpora lutea (per group) - Implantations (per group)] / Corpora lutea (per group) x 100


Post implantation
loss [%] = [Implantations (per group) - living fetuses (per group)] / Implantations (per group) x 100


Calculation of mean indices per litter (see table 7-2 and A7)
Pre implantation
loss [%] = sum of pre-implantation losses per litter in a group [%] / number of litters in a group

Post implantation
loss [%] = sum of post-implantation losses per litter in a group [%] / number of litters in a group



Fetal examinations:
The fetuses were removed and the following examinations performed:
(a) Macroscopic inspection (gross evaluation) of the placentae for example for focal indurations.
(b) The number of fetuses (alive and dead) and placentae (location in uterus and as-signment to the fetus) was determined.
(c) Sex and viability of fetuses were determined. Animals are said to be viable when they are found alive (spontaneous breathing,
spontaneous movement).
(d) Number and size of resorptions were determined.
(e) Corpora lutea in the ovaries, implantations and location of fetuses in the uterus were determined.
(f) Weights of fetuses and weights of the placentae were determined (fetuses were considered as runts if their weight was less than 70% of the mean litter weight).
(g) All fetuses (dead and alive) were inspected externally for damages, especially for malformations .
(h) The fetuses were sacrificed by an ether atmosphere.
(i) Examination of fetuses and determination of number and kind of retardations, variations or malformations:
1) 50% of the number of fetuses in each litter were examined for skeletal anomalies. The thorax and peritoneal cavity (without damage to ribs and sternum) were opened and the location, size and condition of the internal organs were determined.
Then the skeleton was double-stained with Alcian blue for the examination of cartilage and with Alizarin red to reveal ossifications (according to DAWSON). The skeletal system was examined (determination of the number and type of retardations, variations as well as malformations).
2) The remaining 50% of the number of fetuses in each litter were examined for soft tissue anomalies. Body sections were made and examined according to WILSON.
The fetuses were allocated to the evaluation of DAWSON or WILSON on an alternating basis.
Statistics:
Parametrical data:
The statistical evaluation of the parametrical values was done by Provantis using the following settings:
Homogeneity of variances and normality of distribution were tested using the BARTLETT’s and SHAPIRO-WILKS test. In case of heterogeneity and/or non-normality of distribution, stepwise transformation of the values into logarithmic or rank values was performed prior to ANOVA. If the ANOVA yielded a significant effect (p ≤ 0.05), inter-group comparisons with the control group were made by the DUNNETT’s test (p ≤ 0.01 and p ≤ 0.05).

Non-parametrical data
The statistical evaluation of non-parametrical values was done using the FISHER or Chi2 test:
FISHERs exact test, n < 100; (p ≤ 0.05 and p ≤ 0.01)
or
Chi2 test, n ≤ 0.01 (p ≤ 0.05 and p ≤ 0.01)

The respective calculations for the FISHER and Chi2 test were performed using Provan-tis (maternal macroscopic findings at necropsy or findings during the external macro-scopic examination of the fetuses). or an internal computer program (e.g. findings during the fetal skeletal or soft tissue examination).
Note:
The statistical evaluation of the pre- and post-implantation index (per group) using the number of corpora lutea, implantation sites and/ or fetuses per group (see table 7-1 Re-production Data - Summary - Values per Group) was done using StatXact 4.0.1 software, as such a calculation is not possible in Provantis.

Significantly different data are indicated in the summary tables of the result sections of the report.
Indices:
Fertility Index, Viability Index, Resorption Index, Pre-Implantation Loss Index, Post-Implantation Loss Index, Runts Index, Variation Index,
Number of litters having abnormalities, Number of abnormalities per litter
Historical control data:
LPT Background Data, see Appendix 4
Summarized results of the 59 last embryotoxicity studies in Sprague-Dawley rats (Charles River Deutschland GmbH) performed at LPT in the years 2000 to July 2015

Clinical signs:
no effects observed
Description (incidence and severity):
No changes in behavior, the external appearance or the faeces were noted in the control group and in the treatment groups.
Mortality:
no mortality observed
Description (incidence):
No premature deaths were noted in the control group and in the treatment groups (100, 300 or 1000 mg test item/kg b.w./day).
Body weight and weight changes:
no effects observed
Description (incidence and severity):
No test item-related differences in body weight were noted between the dams of the control group and the dams of the treatment groups (100, 300 or 1000 mg test item/kg b.w./day).
Body weight gain
No test item-related changes in body weight gain in comparison to the control group were noted for the dams of the treatment groups (100, 300 or 1000 mg test item/kg b.w./day), when regarding the 3 day intervals of body weight gain, body weight gain after the start of treatment (from gestation day 6 to 21) and body weight gain for the whole study period (gestation day 0 to 21).

No test item-related differences in the body weight at autopsy were noted between the dams of the control group and the dams of the treatment groups (100, 300 or 1000 mg test item/kg b.w./day).
Food consumption and compound intake (if feeding study):
no effects observed
Description (incidence and severity):
No test item-related differences in food consumption were noted between the control group and the treatment groups.
Food efficiency:
not specified
Water consumption and compound intake (if drinking water study):
no effects observed
Description (incidence and severity):
No test item-related changes in drinking water consumption were noted between the dams of the control group and the dams of the treatment groups by visual appraisal.

Ophthalmological findings:
not specified
Haematological findings:
not specified
Clinical biochemistry findings:
not specified
Urinalysis findings:
not specified
Behaviour (functional findings):
not specified
Immunological findings:
not specified
Organ weight findings including organ / body weight ratios:
no effects observed
Description (incidence and severity):
The heart, the liver and the kidneys were weighed and no test item-related differences for the absolute and the relative organ weights were noted between the control group and the treatment groups (100, 300 or 1000 mg test item/kg b.w./day).
Gross pathological findings:
no effects observed
Description (incidence and severity):
No observations were noted for the dams of the control group and the dams of the treatment groups (100, 300 or 1000 mg test item/kg b.w./day) during the macroscopic inspection of the organs and tissues.

Neuropathological findings:
not specified
Histopathological findings: non-neoplastic:
not specified
Histopathological findings: neoplastic:
not specified
Other effects:
no effects observed
Description (incidence and severity):
Gravid uterus and carcass weight
No test item-related differences were noted between the gravid uterus weight and the carcass weight of the control dams and the dams of the treatment groups.

Body weight gain from gestation day 6
No test item-related differences between the control group and the treatment groups were noted for the absolute body weight gain and the net body weight gain (without gravid uterus) between gestation day 6 and gestation day 21 (period after the start of treatment).
Details on results:
no findings
Number of abortions:
no effects observed
Description (incidence and severity):
No influence on the reproductive parameters (number of implantation sites, resorptions and fetuses) was noted, see table below.
Two runts were noted in the control group (no. 3-6 (twin) and no. 10-10) and one runt was noted in the intermediate dose group (no. 66-18).
Pre- and post-implantation loss:
no effects observed
Description (incidence and severity):
No influence on the reproductive parameters (number of implantation sites, resorptions and fetuses) was noted, see table below.
Total litter losses by resorption:
no effects observed
Description (incidence and severity):
No influence on the reproductive parameters (number of implantation sites, resorptions and fetuses) was noted, see table below.
Early or late resorptions:
no effects observed
Description (incidence and severity):
No influence on the reproductive parameters (number of implantation sites, resorptions and fetuses) was noted, see table below.
Dead fetuses:
no effects observed
Description (incidence and severity):
No influence on the reproductive parameters (number of implantation sites, resorptions and fetuses) was noted, see table below.
Changes in pregnancy duration:
not examined
Description (incidence and severity):
Migrated Data from removed field(s)
Field "Effects on pregnancy duration" (Path: ENDPOINT_STUDY_RECORD.DevelopmentalToxicityTeratogenicity.ResultsAndDiscussion.ResultsMaternalAnimals.MaternalDevelopmentalToxicity.EffectsOnPregnancyDuration): not examined
Changes in number of pregnant:
no effects observed
Description (incidence and severity):
No influence on the reproductive parameters (number of implantation sites, resorptions and fetuses) was noted, see table below.
Details on maternal toxic effects:
No test item-related influence on the reproductive parameters (number of implantation sites, fetuses, resorptions and the index of post-implantation) were noted between the dams of the control group and the dams of the treatment groups (100, 300 or 1000 mg test item/kg b.w./day).
see table below "any other information on results"
Key result
Dose descriptor:
NOAEL
Effect level:
> 1 000 mg/kg bw/day
Based on:
test mat.
Basis for effect level:
other: maternal toxicity
Fetal body weight changes:
no effects observed
Description (incidence and severity):
No test item-related differences were noted between the control group and the treatment groups.
Migrated Data from removed field(s)
Field "Fetal/pup body weight changes" (Path: ENDPOINT_STUDY_RECORD.DevelopmentalToxicityTeratogenicity.ResultsAndDiscussion.ResultsFetuses.FetalPupBodyWeightChanges): no effects observed
Field "Description (incidence and severity)" (Path: ENDPOINT_STUDY_RECORD.DevelopmentalToxicityTeratogenicity.ResultsAndDiscussion.ResultsFetuses.DescriptionIncidenceAndSeverityFetalPupBodyWeightChanges): The placental and fetal weights showed no test item-related differences between the control group and the treatment groups.
Reduction in number of live offspring:
no effects observed
Description (incidence and severity):
No dead fetuses were noted in the control group and in the test item-treated groups
Changes in sex ratio:
no effects observed
Description (incidence and severity):
No test item-related differences between the ratio of male and female fetuses were noted between the control group and the treatment groups.
Changes in litter size and weights:
no effects observed
Changes in postnatal survival:
not examined
External malformations:
effects observed, non-treatment-related
Description (incidence and severity):
No macroscopically visible external observations (malformations or variations) were noted for the fetuses of the test item-treated groups during the macroscopic inspection at laparotomy.
However, one fetus (no. 2-3) with an external malformation in the form of a small tail (brachycaudia) was noted in the control group. This finding is in the range of spontaneous variability
Skeletal malformations:
effects observed, non-treatment-related
Description (incidence and severity):
No skeletal malformations were noted for the fetuses of the control group and the test item-treated groups during the skeletal examination according to DAWSON.

Skeletal variations
Skeletal variations were noted for the ribs (wavy or less than 13 rib(s) ossified) and the sternebrae (bipartite, misaligned to a slight degree or misshapen).
No test item-related increase in the incidence of the observed skeletal variations in com-parison to the control group were noted for the fetuses of the treatment groups.

Skeletal retardations
Retardations (delayed ossifications) were related to the skull (incomplete ossification of frontal, parietal, interparietal and/or supraoccipital areas), the hyoid (unossified), the sternum (sternebra(e) incompletely ossified, reduced in size or unossified), the thoracic vertebral bodies (bipartite or dumbbell-shaped), the sacral vertebral bodies (unossified), the caudal vertebral bodies (all bodies unossified), os ischii (incompletely ossified or unossified), os pubis (incompletely ossified) and the metacarpalia (absence of ossification in metacarpalia 2 to 5).
The incidences of the observed skeletal retardations revealed no test item-related differences between the fetuses of the control group and the fetuses of the treatment groups. However, a statistically significantly increased incidence of unossified hyoids was noted at the intermediate dose level. As no dose-response relationship was noted and all values were still in the range of LPT background data, this observation is not considered as test item-related.
Visceral malformations:
effects observed, non-treatment-related
Description (incidence and severity):
The macroscopic inspection of the organs and tissues for gross alterations at laparotomy revealed no malformations or variations for the fetuses of the control group and the fetuses of the treatment groups .

No malformations were noted for the fetuses of the control group and the fetuses of the low and the high dose group during the soft tissue examination according to WILSON.
One fetus with a cleft palate (69-13) was noted at the intermediate dose level. The finding of a cleft palate during the external and/or the soft tissue examination according to WILSON is in the range of LPT back-ground data and not considered as test item-related.

Variations
During the examination of the organs and tissues according to WILSON variations were noted for the cerebral ventricle (dilatation of the 4th ventricle), the kidneys (uni- or bilateral dilatation of the renal pelvis, reduced in size, malpositioned) and the liver (haemorrhagic focus/foci).
No test item-related differences and no statistically significant differences in the incidences of the observed variations were noted between the control group and the test item-treated groups.

Unclassified Observations
A thoracic cavity filled with blood was noted for one fetus of the control group (no. 17-9), one fetus of the intermediate (no. 54-12) and one fetus of the high dose group (no. 78-13). This observation is a preparation induced artefact and not considered as test item-related.

Details on embryotoxic / teratogenic effects:
No test item-related malformation or variations were noted during the macroscopic inspection at laparotomy (including an external inspection and a gross inspection of the organs), the skeletal examination according to DAWSON and the soft tissue examination according to WILSON.
No test item-related retardation (delay in ossification) was noted in any of the treatment groups.
Key result
Dose descriptor:
NOAEL
Effect level:
1 000 mg/kg bw/day (nominal)
Based on:
test mat.
Basis for effect level:
other: developmental toxicity
Key result
Developmental effects observed:
no

Reproduction data of the dams

arameter

Group 1

Control

(n=20)

Group 2

100

mg/kg

(n=20)

Group 3

300 mg/kg

(n=20)

Group 4

1000 mg/kg

(n=20)

 

Corpora lutea

total

mean per dam

299

15.0

290

14.5

310

15.5

294

14.7

 

Implantation sites

total

mean per dam

294 #

14.7

287

14.4

303

15.2

282

14.1

 

Resorptions

total

mean per dam

7

0.4

11

0.6

8

0.4

10

0.5

 

Early resorptions

total

mean per dam

7

0.4

8

0.4

6

0.3

9

0.5

 

Late resorptions

total

mean per dam

0

0.0

3

0.2

2

0.1

1

0.1

 

Live fetuses

total

mean per dam

288

14.4

276

13.8

295

14.8

272

13.6

 

Dead fetuses

total

0

0

0

0

 

Pre-implantation loss [%]

per group ##1

mean per dam

1.7

1.6

1.0

1.4

2.3

2.3

4.1*

3.6

 

Post-implantation loss [%]

per group ##2

mean per dam

2.0

2.3

3.8

4.4

2.6

2.9

3.5

3.7

 

 

*/**:

 p ≤ 0.05/ 0.01

Statistical analyses were performed for the mean values per dam using an ANOVA/DUNNETT test.

 

#

A twin was noted for dam no. 3 for implantation site 6.

 

##1

The statistical comparison of the pre-implantation loss per group was done by comparing the values of implantation sites/corpora lutea of the test group with the ratio of implantation sites/corpora lutea of the control group using the Chi2test.

 

##2

The statistical comparison of the post-implantation loss per group was done by comparing the values of live fetuses/implantation sites of the test group with the ratio of fetuses/implantation sites of the control group using the Chi2test.

 

 

 

Analysis of the test item-formulation

These results indicated correctly prepared and homogenised test item vehicle mixtures, which were stable for at least 24 hours.

The results of the test item-formulation analyses for the investigated parameters are listed in the table below:

Parameter

Sampling / dealing

Range of

nominal concentration

Concentration

immediately after preparation on test days 7 and before administration to the last animal on test day 31

94.6% - 106.0%

Stability

left at room temperature after preparation for 8h or 24 h on test day 7

91.1% - 104.5%

Homogeneity

at start of administration, during administration and before administration to the last animal (on test day 7)

 

99.3% - 107.5%

 

Conclusions:
Under the present test conditions, the no-observed-adverse-effect level (NOAEL) was above 1000 mg test item/kg b.w./day for the dams. None of the dams died prematurely and no signs of clinical toxicity were noted.
The no-observed-adverse-effect level (NOAEL) for the fetal organism was also above 1000 mg test item/kg b.w./day.
The reproductive parameters (number of implantation sites, number of resorptions and number of fetuses) were not influenced by the test item.
No dead fetuses, no malformations and no test item-related variations or retardations were noted.
Under the conditions of the study, test item did not show any teratogenic potential.
Executive summary:

The aim of this dose-range-finding study was the examination of the influence of test item administered orally during the critical period of organogenesis and the fetal development (6th to 20th day of gestation) on the pregnant rat and the fetus.

 

In this prenatal developmental toxicity study, the test item was administered orally to female rats at dose levels of 100, 300 or 1000 mg/kg b.w./day from the 6th to 20th day of pregnancy.

Findings

Examination of the dams:

 

Mortality

No premature deaths were noted in the control group and in the treatment groups

 

Clinical signs

No changes in behavior, the external appearance or the faeces were noted in the control group and in the treatment groups.

 

 

Body weight and

body weight gain

 No test item-related differences in body weight and body weight gain were noted between the control group and the treatment groups.

 

 

Food consumption

 

No test item-related differences in food consumption were noted between the control group and the treatment groups.

 

 

Drinking water consumption

No differences were noted.

 

 

Necropsy findings

No changes were noted during the macroscopic inspection of the dams at necropsy.

 

 

Uterus and carcass weights

No test item-related differences in uterus and carcass weights were noted between the control group and the treatment groups.

 

 

Reproduction data

No influence on the reproductive parameters (number of implantation sites, resorptions and fetuses) was noted.

 

Examination of the fetus

Mortality

No dead fetuses were noted in any of the test groups.

 

Body weight of the fetuses

and the placentae

 

No test item-related differences were noted between the control group and the

treatment groups.

Fetal alterations

 

Malformations

No test item-related malformations were noted during the macroscopic examination at laparotomy (external inspection and inspection of the organs and tissues for gross lesions), the skeletal examination according to DAWSON and the soft tissue examination according to WILSON.

 

 

Variations

The macroscopic examination at laparotomy, the skeletal examination according to DAWSON and the soft tissue examination according to WILSON revealed no test item-related variations.

 

 

Retardations

No test item related retardations were noted during the skeletal examination according to DAWSON.

 

 

Analysis of test item

formulations

The measured actual concentrations of the test item in the test item vehicle-mixtures were between 91.1% and 107.5% of the nominal concentrations, indicating correctly prepared and homogenized formulations which were stable at room temperature for at least 24 h.


Under the present test conditions, the no-observed-adverse-effect level (NOAEL) was above 1000 mg test item/kg b.w./day for the dams.

None of the dams died prematurely and no signs of clinical toxicity were noted.

 

The no-observed-adverse-effect level (NOAEL) for the fetal organism was also above 1000 mg test item/kg b.w./day.

 

The reproductive parameters (number of implantation sites, number of resorptions and number of fetuses) were not influenced by the test item.

 

No dead fetuses, no malformations and no test item-related variations or retardations were noted.

 

Under the conditions of the study, test item did not show any teratogenic potential.

Effect on developmental toxicity: via oral route
Endpoint conclusion:
no adverse effect observed
Dose descriptor:
NOAEL
1 000 mg/kg bw/day
Species:
rat
Quality of whole database:
The study is GLP-compliant and valid without restriction (Klimisch code 1).
Effect on developmental toxicity: via inhalation route
Endpoint conclusion:
no study available
Effect on developmental toxicity: via dermal route
Endpoint conclusion:
no study available
Additional information

A prenatal developmental toxicity (oral) study according to OECD 414 was conducted.

Justification for selection of Effect on developmental toxicity: via oral route:
For Cyclohexane, 5 -isocyanato-1 -(isocyanatomethyl)-1,3,3 -trimethyl-, homopolymer, acetone oxime-blocked a prenatal developmental toxicity (oral) study according to OECD 414 is available.

The no-observed-adverse-effect level (NOAEL) was above 1000 mg test substance/kg b.w./day for the dams.

None of the dams died prematurely and no signs of clinical toxicity were noted.

The no-observed-adverse-effect level (NOAEL) for the fetal organism was also above 1000 mg test substance/kg b.w./day.

Toxicity to reproduction: other studies

Additional information

For Cyclohexane, 5 -isocyanato-1 -(isocyanatomethyl)-1,3,3 -trimethyl-, homopolymer, acetone oxime-blocked a prenatal developmental toxicity (oral) study according to OECD 414 is available.

The no-observed-adverse-effect level (NOAEL) was above 1000 mg testsubstance /kg b.w./dayfor the dams.

None of the dams died prematurely andno signs of clinical toxicitywere noted.

The no-observed-adverse-effect level (NOAEL) for thefetal organismwas also above 1000 mg Test substance/100/kg b.w./day.

Justification for classification or non-classification

Regarding toxicity to reproduction cyclohexane, 5-isocyanato-1-(isocyanatomethyl)-1,3,3-trimethyl-, homopolymer, acetone oxime-blocked, is not classified according to the criteria of EC Directive 67/548/EECand EC Regulation 1272/2008.

Additional information